LOCUS       sequence-297671-        6708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6708)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046325
FEATURES             Location/Qualifiers
     source          1..6708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1303..1320
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2463..2481)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2832..3635
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      3718..4700
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4778..4807
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4829..5416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5428..5455)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5479..5670)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5671..6462,6463..6531))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6294..6313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6532..6634)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6635..6666)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gacttattta tgagaatatt cacgatcggc acagtgaccc taaagcaggg tgagattaaa
      541 gatgctactc catcagattt cgttagggca accgctacta tccccattca agccagcttg
      601 ccttttggtt ggttgattgt aggagtcgct ctcttggctg ttttccaatc cgccagtaag
      661 atcattactt taaaaaagag atggcaattg gcgctttcta agggtgttca cttcgtctgt
      721 aatctgttgt tattgtttgt cactgtttat tcccatttgc tcctagttgc cgctggttta
      781 gaagctccat tcctgtactt gtacgctttg gtctatttcc tgcaatctat caactttgtt
      841 cgtatcatta tgagattgtg gttgtgctgg aaatgtcgtt ctaagaaccc gttgttatac
      901 gacgctaatt acttcctttg ttggcacacc aactgttacg attattgtat tccatataac
      961 tcggttactt cctctatagt tatcacctcc ggggacggta ccacaagtcc aatttcagaa
     1021 catgactacc aaattggtgg ttacactgaa aagtgggagt ctggtgtcaa agattgcgta
     1081 gtgttacaca gctattttac ttctgattac taccagttgt atagcaccca attgtccact
     1141 gacaccggag tggaacacgt cacatttttc atctacaata agatagtcga tgaacctgaa
     1201 gaacatgttc aaatacacac tattgacggc tctagcggtg tagttaaccc agttatggaa
     1261 ccaatctatg atgagccaac aacaacgacc tctgttccct tactcgtgcc gagaggttct
     1321 aaaatcgagg aaggtaagct agtcatttgg ataaatggag ataagggtta caacggtttg
     1381 gcagaagtag gcaagaaatt tgaaaaggac acaggtatca aggtgacggt tgaacatcca
     1441 gacaagttag aagaaaaatt ccctcaagtt gctgctactg gtgatggtcc cgacattatt
     1501 ttctgggccc acgatcgttt tggtggatat gctcagtcag gtttgctggc tgagatcacc
     1561 ccagacaagg ccttccaaga taaattgtac ccattcactt gggatgctgt cagatacaac
     1621 ggtaagctta tagcatatcc aattgctgtt gaagccttat cgttgatcta caataaggac
     1681 ttgttaccta acccaccaaa gacatgggaa gaaattccag ctttggataa agagctaaag
     1741 gccaagggaa aaagcgcttt gatgtttaac ttacaagaac cttatttcac ctggccatta
     1801 atcgctgctg acggtggtta cgcatttaaa tacgaaaacg gtaagtatga cattaaggat
     1861 gtcggcgttg ataatgctgg tgcaaaggct ggtttgacct tcttggttga tttaataaag
     1921 aataaacaca tgaacgctga cactgactac tctatcgccg aagccgcttt caacaagggt
     1981 gaaactgcta tgaccataaa tggtccatgg gcctggagta acattgatac ttctaaagta
     2041 aactatggtg tcactgtttt gcctacattc aaaggccaac catccaagcc ctttgtcggt
     2101 gttctatccg ctggcatcaa tgctgcttct ccaaacaagg agctagctaa agaatttttg
     2161 gaaaattact tgttaactga cgaaggtttg gaggccgtta acaaggacaa gccattgggt
     2221 gctgtcgctt tgaaatctta cgaagaagaa ttggctaagg atcccagaat tgccgctacc
     2281 atggagaacg cccaaaaggg tgaaattatg ccaaacatcc cacaaatgtc agccttctgg
     2341 tacgcagtta ggactgccgt gattaatgct gcttccggta gacaaaccgt cgacgaagca
     2401 ttaaaagatg ctcaaacgta aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     2461 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     2521 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     2581 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     2641 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     2701 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     2761 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     2821 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     2881 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     2941 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     3001 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     3061 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     3121 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     3181 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     3241 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     3301 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     3361 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     3421 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     3481 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     3541 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     3601 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     3661 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     3721 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     3781 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     3841 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     3901 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     3961 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     4021 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     4081 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     4141 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     4201 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     4261 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     4321 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     4381 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     4441 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     4501 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     4561 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     4621 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     4681 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     4741 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     4801 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4861 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4921 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5281 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     5341 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     5401 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     5461 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5521 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5581 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5641 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     5701 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5761 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5821 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     5881 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5941 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6001 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6061 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     6121 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6181 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6241 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6301 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     6361 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6421 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6481 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     6541 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     6601 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     6661 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
